Detection of Phosphorylated and Total ERK and p38 MAPK by Bio-Plex Assay, ELISA, and Western Blotting, Rev A

well and 1 g/well; the reported sample absorbance reached a... ... Conclusions For the detection of... For the detection of phospho-p38 MAPK in HEK 293 cells the Bio-Plex ... For the detection of total ERK the Bio-Plex assay demonstrated bette...

well and 1 g/well;
the reported sample absorbance reached a plateau, and the highest OD reported
was only 1.755 (Figure 4A). The Bio-Plex assay demonstrated distinctly
separate signals across all the samples (Figure 4B), which correlated
well with the western blot bands (Figure 4C).

Conclusions
For the detection of total p38 MAPK in HEK 293 cells, both the Bio-Plex
assay and ELISA correlated well with the western blot. The Bio-Plex assay
showed saturation at the high end but potential to detect the target in
further-diluted samples. With the HeLa lysate, the Bio-Plex assay detected
the target with good sensitivity and demonstrated a broad detection range
that correlated well with the western blot bands. The target was not detectable
by ELISA

For the detection of phospho-p38 MAPK in HEK 293 cells, the Bio-Plex
assay demonstrated a broad detection range at the high end and good correlation
with the western blot. Bio-Plex and ELISA assays both showed potential
for higher sensitivity toward more dilute samples

For the detection of total ERK, the Bio-Plex assay demonstrated better
signal distinction and sensitivity compared to ELISA

For phospho-ERK detection, the ELISA showed saturation at the high
end. The Bio-Plex assay showed better signal distinction, dose response,
and correlation with western blots compared to ELISA

The Bio-Plex suspension array system includes fluorescently labeled microspheres
and instrumentation licensed to Bio-Rad Laboratories, Inc. by the Luminex
Corporation.

Information in this tech note was current as of the date of writing (2004) and
not necessarily the date this version (Rev A, 2004) was published.
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